Cutaneous malignant melanoma is an aggressive cancer of melanocytes with a strong propensity to metastasize. We posit that melanoma cells acquire metastatic capability by adopting an embryonic-like ...phenotype, and that a lineage approach would uncover metastatic melanoma biology. Using a genetically engineered mouse model to generate a rich melanoblast transcriptome dataset, we identify melanoblast-specific genes whose expression contribute to metastatic competence and derive a 43-gene signature that predicts patient survival. We identify a melanoblast gene, KDELR3, whose loss impairs experimental metastasis. In contrast, KDELR1 deficiency enhances metastasis, providing the first example of different disease etiologies within the KDELR-family of retrograde transporters. We show that KDELR3 regulates the metastasis suppressor, KAI1, and report an interaction with the E3 ubiquitin-protein ligase gp78, a regulator of KAI1 degradation. Our work demonstrates that the melanoblast transcriptome can be mined to uncover targetable pathways for melanoma therapy.
Alpha-synuclein is a presynaptic protein that forms abnormal cytoplasmic aggregates in Lewy body disorders. Although nuclear alpha-synuclein localization has been described, its function in the ...nucleus is not well understood. We demonstrate that alpha-synuclein modulates DNA repair. First, alpha-synuclein colocalizes with DNA damage response components within discrete foci in human cells and mouse brain. Removal of alpha-synuclein in human cells leads to increased DNA double-strand break (DSB) levels after bleomycin treatment and a reduced ability to repair these DSBs. Similarly, alpha-synuclein knock-out mice show increased neuronal DSBs that can be rescued by transgenic reintroduction of human alpha-synuclein. Alpha-synuclein binds double-stranded DNA and helps to facilitate the non-homologous end-joining reaction. Using a new, in vivo imaging approach that we developed, we find that serine-129-phosphorylated alpha-synuclein is rapidly recruited to DNA damage sites in living mouse cortex. We find that Lewy inclusion-containing neurons in both mouse model and human-derived patient tissue demonstrate increased DSB levels. Based on these data, we propose a model whereby cytoplasmic aggregation of alpha-synuclein reduces its nuclear levels, increases DSBs, and may contribute to programmed cell death via nuclear loss-of-function. This model could inform development of new treatments for Lewy body disorders by targeting alpha-synuclein-mediated DNA repair mechanisms.
Context. During the period between 15 September 2014 and 4 February 2015, the Rosetta spacecraft accomplished the circular orbit phase around the nucleus of comet 67P/Churyumov-Gerasimenko (67P). The ...Grain Impact Analyzer and Dust Accumulator (GIADA) onboard Rosetta monitored the 67P coma dust environment for the entire period. Aims. We aim to describe the dust spatial distribution in the coma of comet 67P by means of in situ measurements. We determine dynamical and physical properties of cometary dust particles to support the study of the production process and dust environment modification. Methods. We analyzed GIADA data with respect to the observation geometry and heliocentric distance to describe the coma dust spatial distribution of 67P, to monitor its activity, and to retrieve information on active areas present on its nucleus. We combined GIADA detection information with calibration activity to distinguish different types of particles that populate the coma of 67P: compact particles and fluffy porous aggregates. By means of particle dynamical parameters measured by GIADA, we studied the dust acceleration region. Results. GIADA was able to distinguish different types of particles populating the coma of 67P: compact particles and fluffy porous aggregates. Most of the compact particle detections occurred at latitudes and longitudes where the spacecraft was in view of the comet’s neck region of the nucleus, the so-called Hapi region. This resulted in an oscillation of the compact particle abundance with respect to the spacecraft position and a global increase as the comet moved from 3.36 to 2.43 AU heliocentric distance. The speed of these particles, having masses from 10-10 to 10-7 kg, ranged from 0.3 to 12.2 m s-1. The variation of particle mass and speed distribution with respect to the distance from the nucleus gave indications of the dust acceleration region. The influence of solar radiation pressure on micron and submicron particles was studied. The integrated dust mass flux collected from the Sun direction, that is, particles reflected by solar radiation pressure, was three times higher than the flux coming directly from the comet nucleus. The awakening 67P comet shows a strong dust flux anisotropy, confirming what was suggested by on-ground dust coma observations performed in 2008.
Outer thymic cortical large lymphocytes were labeled by transcapsular administration of tritiated thymidine. By 2-4 days after labeling, small and medium labeled lymphocyte descendants were found ...throughout the cortex and in the medulla. The labeled cortical lymphocytes undergo pycnosis after parenteral administration of hydrocortisone 24 h previously, but their (labeled) medullary descendants do not. In addition, parenteral administration of hydrocortisone at the time of surface labeling results in the absence of the appearance of labeled medullary descendants.
CX3CR1 is a chemokine receptor with a single ligand, the membrane-tethered chemokine CX3CL1 (fractalkine). All blood monocytes express CX3CR1, but its levels differ between the main 2 subsets, with ...human CD16+ and murine Gr1low monocytes being CX3CR1hi. Here, we report that absence of either CX3CR1 or CX3CL1 results in a significant reduction of Gr1low blood monocyte levels under both steady-state and inflammatory conditions. Introduction of a Bcl2 transgene restored the wild-type phenotype, suggesting that the CX3C axis provides an essential survival signal. Supporting this notion, we show that CX3CL1 specifically rescues cultured human monocytes from induced cell death. Human CX3CR1 gene polymorphisms are risk factors for atherosclerosis and mice deficient for the CX3C receptor or ligand are relatively protected from atherosclerosis development. However, the mechanistic role of CX3CR1 in atherogenesis remains unclear. Here, we show that enforced survival of monocytes and plaque-resident phagocytes, including foam cells, restored atherogenesis in CX3CR1-deficent mice. The fact that CX3CL1-CX3CR1 interactions confer an essential survival signal, whose absence leads to increased death of monocytes and/or foam cells, might provide a mechanistic explanation for the role of the CX3C chemokine family in atherogenesis.
Much of the differentiation of murine T cells takes place in the thymus, perhaps influenced by the operation of stringent selection mechanisms whose existence has been inferred from the high rate of ...thymocyte turnover in the absence of extensive emigration. The origin of those 1% of total thymocytes which leave the thymus and seed the peripheral lymphoid organs is obscure. Recent thymic emigrants are functionally and phenotypically mature, and the purported greater maturity of medullary relative to cortical thymocytes is often cited a evidence for the medullary origin of thymic emigrants, a suggestion not without its critics. To approach this question, we have now isolated a a subpopulation of thymocytes expressing high levels of a receptor that mediates the homing of blood-borne lymphocytes into peripheral lymph nodes. Surprisingly, this population of cells (1-3% of total thymocytes) is both cortical and immunocompetent, containing approximately half of all thymic cytolytic T-lymphocyte precursors. The combination of homing receptor expression and immunocompetence makes this cortical population ideally suited for emigration to peripheral lymphoid organs.
We have previously demonstrated that in vitro cell lines of mouse thymic lymphomas express surface receptors specific for the retrovirus that induced them. This study extends these observations to an ...analysis of receptor-bearing cells in the preleukemic and leukemic phases of spontaneous AKR thymic lymphomagenesis. AKR mice regularly begin expressing N-tropic retroviruses (as assayed on NIH fibroblasts by the XC plaque assay) in several tissues early in life; thymic lymphocytes also express these viruses, but are not autonomously transformed. Later thymic lymphomas emerge which are capable of metastasizing in the host of origin or transplanting leukemias into syngeneic hosts. Just prior to the appearance of thymic lymphomas, these mice also begin producing xenotropic retroviruses as assayed in xenogeneic (For example, mink) fibroblasts, and concomitant with the appearance of the leukemias is the appearance of "recombinant" retroviruses which cause mink fibroblast foci (MCF); these viruses express elements of both N- and X-tropic virus envelopes and N-tropic viral gene products in their cores. Spontaneous AKR leukemias also produce other retroviruses which do not cause XC plaques or mink fibroblast foci; these are called SL viruses. The subject of this study was to test whether in vivo thymocytes in the preleukemic and leukemic periods also bear receptors specific for N-tropic, recombinant MCF and SL AKR retroviruses. We demonstrated that each spontaneous thymic lymphoma does bear receptors that bind viruses produced by the lymphomas and MCF-247 to a high degree and that bind N-ecotropic AKR retroviruses less well. Thymic lymphocytes predominating in the preleukemic period do not express detectable levels of receptors for either of the viruses. In some mice, receptor-positive cells co-exist with receptor-negative cells; only the receptor-positive cells are capable of transplanting leukemia to syngeneic hosts. We conclude that the presence of specific cell surface receptors for lymphoma cell-produced and recombinant AKR retroviruses is a marker for leukemia in these hosts.
Cancer immunotherapy has emerged as a promising therapeutic intervention. However, complete and durable responses are only seen in a fraction of patients who have cancer. A key factor that limits ...therapeutic success is the infiltration of tumors by cells of the myeloid lineage. The inhibitory receptor signal regulatory protein-α (SIRPα) is a myeloid-specific immune checkpoint that engages the “don’t eat me” signal CD47 expressed on tumors and normal tissues. We therefore developed the monoclonal antibody KWAR23, which binds human SIRPα with high affinity and disrupts its binding to CD47. Administered by itself, KWAR23 is inert, but given in combination with tumor-opsonizing monoclonal antibodies, KWAR23 greatly augments myeloid cell-dependent killing of a collection of hematopoietic and nonhematopoietic human tumor-derived cell lines. Following KWAR23 antibody treatment in a human SIRPA knockin mouse model, both neutrophils and macrophages infiltrate a human Burkitt’s lymphoma xenograft and inhibit tumor growth, generating complete responses in the majority of treated animals. We further demonstrate that a bispecific anti-CD70/SIRPα antibody outperforms individually delivered antibodies in specific types of cancers. These studies demonstrate that SIRPα blockade induces potent antitumor activity by targeting multiple myeloid cell subsets that frequently infiltrate tumors. Thus, KWAR23 represents a promising candidate for combination therapy.
Cells staining for Lyt-1 are more frequent than cells staining for Lyt-2 in both primary follicles and the cuffs of secondary follicles; there is an even more striking predominance of cells bearing ...only Lyt-1 in germinal centers. In addition, there is an increase in the total percentage of cells bearing T cell antigens in germinal centers compared to primary follicles. These differences in phenotype and distribution of T cell populations indicate the T cells in B cell areas, and especially in germinal centers, are not randomly selected, but rather represent a specific subpopulation of T cells enriched for the helper phenotype (Lyt-1+2-), perhaps involved in the development and/or function of germinal centers.
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